| 名稱 | PF 477736 |
| 描述 | PF 477736 (PF-736,PF-00477736) is a specifc, effective and ATP-competitive Chk1 inhibitor (Ki: 0.49 nM ) and also inhibits FGFR3, Aurora-A, VEGFR2, Flt3, Fms (CSF1R), Ret and Yes. |
| 細胞實驗 | The IC50 assay measures the antiproliferative effects of PF-477736 on p53-defective human cancer cell lines. Cells in each line are seeded in complete medium at an exponentially growing density in 96-well assay plate and allowed to attach for 16 hours. Serial dilutions of PF-477736 are then done, and appropriate controls are added to each plate. Cells are incubated with drug for 96 hours. After incubation, MTT working stock diluted in complete medium is added to each well, and cells are incubated for 4 hours. After centrifugation and supernatant removal, DMSO is added to each well and plates are read on SpectraMax plate reader at 540 nm. (Only for Reference) |
| 激酶實驗 | Binding assay: The assay is performed in a 96-well plate for 20 minutes at 30℃ in 0.1 mL of assay buffer containing 50 mM TRIS pH 7.5, 0.4 M NaCl, 4 mM PEP, 0.15 mM NADH, 28 units of lactate dehydrogenase/mL, 16 units of pyruvate kinase/mL, 3 mM DTT, 0.125 mM Syntide-2, 0.15 mM ATP and 25 mM magnesium chloride. Assays are initiated with 1 nM of CHK1 kinase domain. The inhibition of CHK1 activity is determined by measuring initial velocities in the presence of varying concentrations of PF-477736. The data is analyzed using Enzyme Kinetic and Excel software and fit to a kinetic model for competitive inhibition to obtain a Ki value. The kinase selectivity of PF-477736 is evaluated by screening the compound at 1 μM or 10 μM against a panel 2 of about 100 protein kinases. |
| 體外活性 | PF-477736 (128 nM) 以劑量依賴性方式消除了CA46及HeLa細胞中由喜樹堿引起的DNA損傷檢查點。PF-477736 在HT29細胞中有效地消除了由吉西他濱引起的S期阻滯，并相應增加了凋亡細胞群體。PF-477736 (540 nM) 以時間和劑量依賴的方式增強了HT29細胞中吉西他濱引起的細胞毒性。PF-477736 增強了一系列化療化合物在MTT檢測中對廣泛p53缺陷的人類癌癥細胞系的生長抑制活性。PF-477736 (360 nM) 加入到吉西他濱停滯的細胞中，引起H2AX磷酸化強度的顯著增加，反映了在DNA損傷附近γ-H2AX分子數(shù)量的增加。PF-477736 (0.5 nM) 在HL-60細胞中存在姜黃素時選擇性阻斷p73和P53的磷酸化。PF-477736 (360 nM) 抑制了COLO205細胞中由多西他賽引起的組蛋白H3 (Ser10) 和Cdc25C (Ser216) 的磷酸化，并增強了凋亡。PF-477736 (250 nM) 與MK-1775聯(lián)合使用在OVCAR-5細胞中顯示出顯著的協(xié)同細胞毒作用，并導致OVCAR-5細胞中含有2N至4N之間DNA含量的細胞積累，以及DNA損傷導致的凋亡細胞死亡。 |
| 體內(nèi)活性 | PF-477736 (4 mg/kg i.v.) 在大鼠體內(nèi)的半衰期（T1/2）為2.9小時，AUC為5.72 μg×hr/mL，CLp為11.8 mL/min/kg。PF-477736 劑量依賴性地增強了在Colo205異種移植小鼠模型中最大耐受劑量的gemcitabine的抗腫瘤活性。PF-477736 (12 mg/kg) 在Colo205異種移植小鼠模型中誘導了組蛋白H3（Ser10）和磷酸化組蛋白H2AX的磷酸化增加。[1] PF-477736 (15 mg/kg i.p.) 在COLO205和MDA-MB-231異種移植模型中增強了docetaxel誘導的腫瘤生長抑制和腫瘤生長延遲。[3] PF 477736（10 mg/kg 每天一次 i.p.）與MK-1775（30 mg/kg 每天兩次口服）聯(lián)用在攜帶OVCAR-5異種移植物的小鼠中實現(xiàn)了更大的腫瘤生長抑制。[4] |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year
Shipping with blue ice/Shipping at ambient temperature. |
| 溶解度 | DMSO : 50 mg/mL (119.2 mM), Sonication is recommended.
10% DMSO+40% PEG300+5% Tween 80+45% Saline : 5 mg/mL (11.92 mM), Solution.
10% DMSO+90% Saline : < 5 mg/mL (11.92 mM), Lower concentrations may be soluble, but exact solubility limit is unknown.
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| 關鍵字 | Yes | VEGFR2 | VEGFR | Src | RET | PF-736,PF00477736 | PF-736,PF 00477736 | PF-00477736 | PF00477736 | PF 477736 | FLT3 | FGFR | cRET | Chk2 | Chk1 | Checkpoint Kinase (Chk) | c-Fms | cFms | CDK | AuroraKinase | Aurora Kinase |
| 相關產(chǎn)品 | 2-Chloropyrazine | Amlexanox | Ribociclib | Lenvatinib | Chloramphenicol | Abemaciclib | Ferulic Acid | Pazopanib | Dasatinib | Sorafenib | Ibrutinib | Kojic acid |
| 相關庫 | 抑制劑庫 | 經(jīng)典已知活性庫 | 抗癌活性化合物庫 | 已知活性化合物庫 | 激酶抑制劑庫 | 抗衰老化合物庫 | 膜蛋白靶向化合物庫 | 藥物功能重定位化合物庫 | 酪氨酸激酶分子庫 | 抗癌臨床化合物庫 | 抗癌藥物庫 | 細胞重編程化合物庫 |